摘要
[目的]观察电针人中穴(GV26)对脑梗死(CI)后内源性神经干细胞增殖、分化的影响。[方法]Wistar大鼠随机分为电针组、模型组、假手术组、空白组,前3组每组24只,按干预和取材的时间分为1、3、7、14 d 4个亚组,空白组6只,采用神经功能缺损体征评分(NSS)评定各组大鼠神经功能缺损程度;苏木精-伊红(HE)染色法观察CI后内源性神经干细胞的形态改变;免疫组化、免疫荧光双标记法检测CI后内源性神经干细胞的增殖、分化。[结果]神经功能方面,空白组、假手术组NSS评分无变化,神经功能正常,模型组、电针组CI大鼠随着时间的延长,两组大鼠在感觉、反射、肢体功能等方面均有改善,3、7、14 d时电针组NSS评分低于模型组,两组差异有统计学意义(P<0.05);HE染色,空白组和假手术组脑组织形态未见明显异常,模型组和电针组均可见神经细胞的细胞核固缩、伴细胞空泡,随干预天数增加,逐渐有结缔组织、星形胶质细胞与正常细胞生成,电针组整体修复情况优于模型组;免疫组化,与空白组、假手术组比较,模型组大鼠各时相内源性神经干细胞(eNSPCs)特异性标识物齿状回巢蛋白(Nestin)目标蛋白平均光密度值(AOD)、胶质纤维酸性蛋白(GFAP)AOD均明显升高(P<0.01),与模型组比较,电针组各时相Nestin AOD均较高(P<0.05或P<0.01),3、7、14 d时GFAP AOD均升高(P<0.05或P<0.01);免疫荧光双标记,与空白组、假手术组比较,模型组大鼠Nestin及GFAP表达量均明显升高(P<0.01),荧光信号强,与模型组比较,电针组各时相Nestin表达均升高(P<0.05或P<0.01),Nestin/5-溴脱氧尿嘧啶核苷(BrdU)共标记信号强,3、7、14 d时GFAP表达升高(P<0.05或P<0.01),GFAP/BrdU共标记信号强。[结论]电针人中穴可明显改善CI大鼠神经功能损伤情况,促进内源性神经干细胞的增殖、分化为星形胶质细胞,从而有利于CI后神经功能网络的修复和功能的重建。
[Objective]The effect of renzhong(GV26)electroacupuncture on the proliferation and differentiation of endogenous neural stem cells after cerebral infarction(CI)was observed.[Methods]Wistar rats were randomly divided into electroacupuncture group,model group,sham surgery group and blank group,the first three groups were 24 in each group,divided into 4 subgroups according to 1,3,7 and 14 d,and 6 rats in blank group,and the degree of neurological deficit in each group was assessed by NSS.HE staining was used to observe the morphological changes of endogenous neural stem cells after CI.Immunohistochemistry and immunofluorescence double labeling to detect the proliferation and differentiation of endogenous neural stem cells after CI.[Results]In terms of neurological function,there was no change in the NSS score of the blank group and the sham surgery group;the neurological function was normal,and the CI rats in the model group and the electroacupuncture group improved their sensation,reflex and limb function with the extension of time,and the NSS scores of the electroacupuncture group at 3,7 and 14 d were lower than those in the model group,and the difference between the two groups was statistically significant(P<0.05);HE staining,blank group and sham surgery group showed no obvious abnormalities in brain tissue morphology,the model group and electroacupuncture group showed nerve cell nucleus contraction,with cell vacuoles,with the increase of intervention days,gradually connective tissue,astrocytes and normal cell production,the overall repair of electroacupuncture group was better than that of the model group.Compared with the blank group and sham group,Nestin AOD and GFAP AOD were significantly increased(P<0.01)in each time phase of rats in the model group,and Nestin AOD was higher(P<0.05 or P<0.01)in the electroacupuncture group compared with the model group,and GFAP AOD was increased(P<0.05 or P<0.01)at 3,7 and 14 d.Compared with the blank group and sham group,the expression of Nestin and GFAP in rats in the model group was significantly increased(P<0.01)and the fluorescence signal was strong,compared with the model group;the expression of Nestin in each phase of the electroacupuncture group was increased(P<0.05 or P<0.01);the Nestin/BrdU co-labeling signal was strong,the GFAP expression was elevated(P<0.05 or P<0.01)at 3,7 and 14 d,and the GFAP/BrdU co-labeling signal was strong.[Conclusion]Renzhong electroacupuncture can significantly improve the neurological damage of CI rats,promote the proliferation and differentiation of endogenous neural stem cells into astrocytes,and thus facilitate the repair and functional reconstruction of neural functional networks after CI.
作者
殷秀梅
杨丽红
韩佳炜
陈林玲
秋添
姜涛
杜元灏
YIN Xiumei;YANG Lihong;HAN Jiawei;CHEN Linling;QUI Tian;JIANG Tao;DU Yuanhao(First Teaching Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300381,China;National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion,Tianjin 300381,China)
出处
《天津中医药》
CAS
2023年第3期325-333,共9页
Tianjin Journal of Traditional Chinese Medicine
基金
天津市教委科研计划项目(2018KJ025)。
关键词
脑梗死
电针
人中穴
内源性神经干细胞
神经功能网络
cerebral infarction
electroacupuncture
renzhong(GV26)
endogenous neural stem cell
neurofunctional networks